Amino-allyl Reverse Transcription

(this protocol was adapted by Joe DeRisi from one developed at Rosetta
Inpharmatics)

Resuspend the contents of one pack of the monofunctional NHS-ester Cy3 or Cy5
dye (mono-functional Cy3 or Cy5 reactive pack, Amersham catalog PA 23001 and PA
25001) in 72 ul DMSO. Aliquot 16 x 4.5 ul and either use immediately or dry the
aliquots in a speed vac. Store aliquots at 4 C in a dessicator. Use one aliquot
of the dyes for each RT labelling reaction and microarray hybridization.

III. CleanupTo continue with the amino-allyl dye coupling procedure all Tris must
be removed from the reaction to prevent the monofunctional NHS-ester Cye
dyes from coupling to free amine groups in solution.